Research Paper Volume 15, Issue 10 pp 4498—4509
Expression of αA-crystallin (CRYAA) in vivo and in vitro models of age-related cataract and the effect of its silencing on HLEB3 cells
- 1 Ningxia Medical University Key Laboratory of Ningxia Minority Medicine Modernization Ministry of Education, Ningxia Medical University, Yinchuan 750004, Ningxia, China
- 2 School of Traditional Chinese Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia, China
- 3 School of Clinical Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia, China
- 4 School of Pharmacy, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
Received: March 4, 2023 Accepted: May 16, 2023 Published: May 28, 2023
https://doi.org/10.18632/aging.204754How to Cite
Copyright: © 2023 Ma et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Aim: To investigate the expression of αA-crystallin (CRYAA) in age-related cataract (ARC) models and its role in lens epithelial cells (LECs).Methods: We used Flow cytometry to detect the apoptosis and cell cycle in HLEB3 cells and Real-time fluorescence quantitative polymerase chain reaction to detect the expression of CRYAA mRNA in HLEB3 and in rabbit lens. The expression of CRYAA in HLEB3 cells and rabbit lenses as well as the proteins related to apoptosis and autophagy in transfected cells were detected by western blotting. The lens structure in rabbits was investigated using hematoxylin-eosin staining. Protein thermostability assay was performed to detect the thermal stability of rabbit lens proteins. CCK- 8 assay was used to detect the viability of transfected cells, and the transfection was recorded by fluorescence photography.
Results: Hydrogen peroxide can promote apoptosis and arrest the cell cycle in HLEB3 cells, and naphthalene can cause cataract formation and damage the structure of the lens in rabbits. Both ARC models can reduce the expression of CRYAA. The expression of CRYAA silencing increased apoptosis and autophagy in HLEB3 cells.
Abbreviations
ARC: Age-related cataract; LECs: lens epithelial cells; H2O2: hydrogen peroxide; CRYAA: αA-crystallin; CLA: chaperone-like activity; CCK8: Cell Counting Kit-8; WB: Western Blotting.